- Fahimeh Kazemi added an answer:How do you transfuse blood in mice?
From what I was reading, the procedure involves euthanizing the donor mouse and collecting blood (Intracardiac) in a heparinized tube and giving the blood via tail vein within the hour. Does anyone have experience with this and possibly have a precise protocol in your lab or your institution on specific steps?
Cardiac puncture is a suitable technique to obtain a single, large, good quality sample from a euthanized mouse or a mouse under deep terminal anaesthesia if coagulation parameters, a separate arterial or venous sample or cardiac histology are not required. It is appropriate for all strains of mouse (euthanizing mouse by the intraperitoneal (i.p.) injection of pentobarbital sodium (100 mg/kg body weight)
0.1 - 1 ml of blood can be obtained depending on the size of the mouse and whether the heart is beating. Blood samples are taken from the heart, preferably the ventricle, which can be accessed either via the left side of the chest, through the diaphragm, from the top of the sternum or by performing a thoracotomy. Blood should be withdrawn slowly to prevent the heart collapsing.
Cardiac puncture should not be used if the peritoneum needs to be lavaged to harvest cells, as this technique can cause blood to escape into the peritoneal cavity.
Cardiac Puncture Blood Collection (Terminal Procedure)
• Mouse: 23-25 gauge needle and 1-3 ml syringe
• Rat: 20-25 gauge needle and 10-20 ml syringe
1. Anesthetize animal. (Surgical plane of anesthesia is required!)
2. Test for reaction by corneal reflex and toe pinch.
3. Blood may be obtained through a ventral, left lateral, or open approach.
4. Ventral approach (closed)
a. Place animal on back (dorsal recumbency)
b. Palpate heart
c. Insert needle slightly left of and under sternum, directed toward animal’s head.
d. Needle and syringe should be held 20-30 degrees off horizontal.
e. Insert into heart
5. Left Lateral approach (closed)
a. Place animal on right side (right lateral recumbency)
b. Palpate heart on left lateral thoracic wall (approximately at point of flexed elbows, between ribs 5 and 6)
c. Insert needle slowly (between ribs and perpendicular to the body) and into heart
6. Open approach
a. Place animal on back (dorsal recumbency)
b. Wet skin on the abdomen with 70% alcohol
c. Make a V-cut through the skin and abdominal wall ~1cm caudal to the last rib
d. Move internal organs to the side
e. Insert needle through the diaphragm and into the vena cava or heart
7. Gently apply negative pressure on syringe plunger. Heart chamber may collapse if negative pressure is too great.
8. Never move the needle side-to-side as this could lacerate the heart or vena cava!
9. If no blood appears, slowly withdraw the needle so that it remains just under the skin or the diaphragm and redirect in a slightly different direction.
10. If blood stops flowing, slowly rotate needle or move it slightly in or out.
11. Withdraw needle after blood has been collected.
12. Perform secondary method of euthanasia to ensure that animal is deceased.
Blood Volumes Expected at Exsanguinations (~3% of Body Weight):
• 25 g Mouse: ~0.75 ml
• 300 g Rat: ~9.0 ml
Blood collects directly from the heart in EDTA or heparin containing tubes, separates by centrifugation (at 3000 g for 10 min at at 4 °C), frizzes, and stores at -80 °C until biochemical analysis was performed.
- Giuliano Grazzini added an answer:Is anyone familiar with the use of Umbilical Cord Blood (UCB) for transfusion?
Can we use umbilical cord blood for transfusion purposes in human? If somebody has any experiences, please share. It would be nice if somebody could share some studies related to it. Thanks!
And also this one:
Allogeneic cord blood red cells for transfusion.
Bianchi M, Landini A, Giannatonio C, Papacci P, d'Onofrio G, Zini G.
Transfus Med Rev. 2012 Jan;26(1):90-1; author reply 91-2. doi: 10.1016/j.tmrv.2011.06.002. Epub 2011 Aug 9. No abstract available.
- Kailash C. Agarwal added an answer:Does anyone have experience with still disease and febrile non-hemolytic reaction?How can one manage fever in adult with still disease and during a blood transfusion? The fever was not present at the beginning of the transfusion.
Not of my expertise. Sorry.Following
- Mojtaba Akhtari added an answer:Does blood transfusion increase the risk of GVHD in patients who are post hematopoietic stem cell transplantation?
Is there any evidence that blood transfusion increase the risk of GVHD in patients post hematopoietic stem cell transplantation?
Transfusion-associated GVHD is seen in patients who receive RBC transfusion and they develop skin rash and pancytopenia. In those patients the bone marrow gets hit and it is almost always fatal, and steroids do not work. However, in GVHD after allogeneic HSCT the hematopoiesis in the bone marrow is from the donor; so, it does not get hit by the GVHD process. We use steroids and they are usually effective. Attached you find a retrospective study about the relationship between RBC transfusion and GVHD from the BMT group at Emory University which is quite interesting, it was presented at ASH last December.Following
- Gregory Barshtein added an answer:Can anybody advise us or provide us with the InSpectra StO2 Monitor (650) and/ or the Aimago Laser-Doppler Imager (Easy LDI)?
For our research on blood transfusion and the micro-circulation, we are seeking to borrow or purchase used InSpectra StO2 Monitor (650) and the Aimago Laser-Doppler Imager (Easy LDI). We will be glad as well to receive the instruments for collaborative research from interested researchers.
Irina & Josef, many thanks for the tip.Following
- Terry S. Singeltary added an answer:Does any country or region consider the introduction of prion filter or bi-functional (prion and leukocyte) filter for blood transfusion?Four cases of transmission of variant CJD in humans from transfused blood components have been reported. Recent investigations have demonstrated the utility of prion-filter and bi-functional (prion+leukocyte) filter with reduction of 3 to 4 log infectivity titer. I am inquiring whether any country or region blood service is considering the adoption of any prion-reduction filter for the prevention of transmission of vCJD. Furthermore, if not, the reason why not, cost, lacking evidence of effectiveness, or concerns for safety?
Wednesday, July 23, 2014
After the storm? UK blood safety and the risk of variant Creutzfeldt-Jakob Disease
22. SaBTO’s decision not to recommend the adoption of prion filtration, taken alongside the other evidence that we have gathered during this inquiry, in our view signals a change from what was a genuinely precautionary approach to vCJD risk reduction in the late 1990s to a far more relaxed approach today. Much of the uncertainty surrounding prions, their potential modes of transmission and the possible rate of undetected infection and disease remains: recent evidence that subclinical prevalence could be as high as one in 2,000 people would suggest that a precautionary approach is now more warranted than ever. (Paragraph 94)
23. Our fear is that the Government’s current attitude is driven less by the available scientific evidence than it is by optimism: a hope that the storm has now passed and that vCJD is no longer the threat to public health that it once was. In the current economic environment, this attitude is not surprising. However, it is not justified. For all we know, the storm may well be ongoing. We conclude this report by recommending that the Government take a more precautionary approach to both vCJD risk mitigation and blood safety more generally, in order to safeguard against future infections. We suggest that it begin by assessing the key risks, known and unknown, that the UK blood supply currently faces and might face in the future, so that it can identify and fill relevant knowledge gaps and support the development of appropriate risk reduction measures and technologies. The Government should initiate this work immediately and we ask that it provide us with an update on its progress well before the dissolution of Parliament. (Paragraph 95)
56After the storm? UK blood safety and the risk of variant Creutzfeldt-Jakob Disease EMBARGOED ADVANCE COPY Not to be published in full, or in part, in any form before 00.01am on Thursday 24 July 2014 Formal Minutes Wednesday 16 July 2014
Friday, October 11, 2013
Removal of exogenous prion infectivity in leukoreduced red blood cells unit by a specific filter designed for human transfusion
Thursday, January 22, 2015
Transmission properties of atypical Creutzfeldt-Jakob disease: a clue to disease etiology?
MAD COW DISEASE nvCJD TEXAS CASE NOT LINKED TO EUROPEAN TRAVEL CDC ***
Sunday, November 23, 2014
*** Confirmed Variant Creutzfeldt-Jakob Disease (variant CJD) Case in Texas in June 2014 confirmed as USA case NOT European ***
the patient had resided in Kuwait, Russia and Lebanon. The completed investigation did not support the patient's having had extended travel to European countries, including the United Kingdom, or travel to Saudi Arabia. The specific overseas country where this patient’s infection occurred is less clear largely because the investigation did not definitely link him to a country where other known vCJD cases likely had been infected.
Sunday, December 14, 2014
*** ALERT new variant Creutzfeldt Jakob Disease nvCJD or vCJD, sporadic CJD strains, TSE prion aka Mad Cow Disease United States of America Update December 14, 2014 Report ***
kindest regards, terryFollowing
- Steingrimur Stefansson added an answer:What are the blood substitutes available for transfusion?
Natural blood has problems of shelf life and infections such as HIV , Hbs Ag.
Also, Jehovah's Witnesses do not understand hematology and biochemistry.
All of the proteins and cells in your blood have a finite lifetime. After a period, they are absorbed by the liver or kidneys, cleaved to their individual components and recycled.
This happens in all of us, every day
Blood transfusion have nothing to do with consumption of human flesh.
Blood transfusions administered to trauma victims are not designed to keep them well fed on human flesh, they are done to keep them alive by replacing their lost blood.
If you are a vampire, then yes, blood is a consumable. But for the rest of us, blood transfusions are procedures that have saved many more lives than prayers.Following
- Niels Lion added an answer:Do you use a platelet additive solution for platelets?
1. Yes, or no? 2. If yes, how many percentage of platelet components? 3. Name of the solution? 4. Are there any solution-associated problems?
1. Yes we use PAS for all platelet concentrates (both apheresis and buffy-caot derived). We use Intersol in a ratio of 39% plasma / 61% Intersol. This ratio comes from the Intercept PI technique specification (32-47% plasma; whereas 100% plasma is feasible, it poses other operational constraints - longer procedure basically).
2. 100% of platelet components. For apheresis we use 500 mL Intersol bags with automatic addition of Intersol on Trimas. For Buffy coat platelets, we use 280 mL Intersol bags, automatically added on the Orbisac machine during pooling of BC.
3. The transition from 100% plasma to 39% plasma /61% Intersol for paltelet storage has been associated in Switzerland with a significant decrease of transfusion reactions:
Whereas the document compares pre- and post Intercept (pathogen inactivation technique) introduction, the main difference for non-infectious transfusion reactions comes from the use of additive solution instead of 100% plasma.
- Mehran Ghasemzadeh added an answer:Why are platelet pockets stored at room temperature (20-25°C)?
Platelet pockets intended for transfusion are stored at room temperature. Is there any risk of protein degradation that could affect their role?
AS main problem, cold storage of platelets leads to clustering of alpha subunits of glycoprotein Ib (GPIbalpha)) on the platelet surface, its Desialylation that accelerating platelet clearance as well as increased rate of apoptosis.
- Khalid A. Altirkawi added an answer:Do you subscribe to the practice of withholding feeding in preterm infants when they receive a blood transfusion?The association of necrotizing enterocolitis (NEC) and blood transfusion is suggested by several case series and observational studies. The few RCTs dealt with this matter so far have not confirmed such association. Nonetheless, many colleagues like to err on the conservative side, i.e. to stop feeding altogether upon blood transfusion. Do you support this approach in your practice?
Two recently published studies are probably of interest in this discussion.
1- Packed red blood cell transfusion is not associated with increased risk of necrotizing enterocolitis in premature infants. by R Sharma, D F Kraemer, R M Torrazza, V Mai, J Neu, J J Shuster, M L Hudak.
2- Red Blood Cell Transfusion Is Not Associated with Necrotizing Enterocolitis: A Review of Consecutive Transfusions in a Tertiary Neonatal Intensive Care Unit. by Matthew B. Wallenstein, Yassar H. Arain, Krista L. Birnie, Jennifer Andrews, Jonathan P. Palma, William E. Benitz, Valerie Y. Chock.
- Blood transfusion in acute MI with multivessel disease the right course for patient?
this is somewhat of a hot topic. 75 y.o. male, BMI 27, comes in for a STEMI with cardiac arrest, primary PCI of prox LAD (2 overlapping DES, radial approach) in 3-VD, ICU, LV EF 25%.
On day 5 he undergoes completion of revascularization with 2 DCBs in the Circ, and 1 DES in the RCA (radial approach).
Hemoglobin begins with 14.5 g/dl on admission and then drops progressively to 12, 10, and then, after the second PCI to 7.5 g/dl, without signs of overt bleeding except for an already known hemorragic gastritis.
Patient currently suffers heart failure.
Should I go for blood transfusion now, or not?
A tu disposición!Following
- What is your experience with Patient Blood Management (PBM) programs in your country? I would like to know whether PBM is a topic in your country. If yes, then which professions are the key drivers of the PBM project? Do you have results from these PBM Projects? Are they safe? Are they effective? What are clinical outcomes?
Thank you very much for sharing the Northern Ireland experience with us. From the view of hemotherapy, the most important benefit of improvements in management of preoperative anemia and perioperative patient care is the increase in overall patient safety
I think, you made an important point by adding high quality audits, which in our hands also help to increase the quality of hemotherapy.
- Mahdi Najafi added an answer:Do you follow blood management guidelines in your daily practice?
Do you follow blood conservation/transfusion guidelines in perioperative care in cardiac surgery? If so, do you use known international/regional guidelines or an institutional protocol?
What are important points in the development and/or implementation of an efficient protocol for blood conservation/transfusion?
Recently a conference was held in our hospital and experts from different disciplines discussed on challenging aspects of coagulation control and transfusion. We had a panel on prerequisites of an appropriate protocol for our hospital. We came to agreement on some topics:
1. To determine time interval from discontinuing anticoagulants and surgery
2. To rely more on TEG results for bleeding control
3. To agree on fibrinogen indications and dosage
4. To measure heparin level (Anti X) in bleeding patients
5. To agree on the indications, route of use and dosage of vitamin K in patients on warfarin who are candidates for emergent surgery
To agree on the critical Hb level for transfusion in different situations.
However, to be honest there is a deep valley between knowledge and practice.Following
- Does anyone have experience with non-invasive Hb measurement in blood donors? Hemoglobin measurement in healthy volunteer blood donors is mandatory before whole blood donation in a great number of countries. However, the current invasive methods are not very reliable. Has anyone broad experience with the non-invasive techniques in daily use?
Thank you very much for sharing your very important data.
- How do you deal with the West-Nile-Virus (WNV) threat in Europe re. blood donors?
In central Europe, West-Nile-Virus (WNV) infections are endemic in small regions and only seasonal small outbreaks occur outside these regions. Therefore, WNV infection risk for blood donors is difficult to assess. One can test all donors by WNV PCR, but that is extremely costly and far more than 99% will be tested negative. However, in the U.S. with their high prevalence of WNV, the WNV PCR has proven cost effective. Alternatively, European blood transfusion centers defer blood donors travelling through "WNV regions" for 4 weeks. What is the strategy, you and your Institution prefers?
Thank you very much. Most useful.
- Bahri Abayli added an answer:What are the indications for transfusion of fresh frozen plasma in a cirrhotic patient?Patient, 55 years old, diaphragm hernia operation.
Cirrhosis is detected intraoperatively (no clinical manifestations). This patient doesn't show history of viral infection of the liver or the use of toxic substances.
The only laboratory parameter which was without normal range: prothrombin 28% with INR 2.36, in postoperative period.
The patient shows no signs of active bleeding, the response to administration of vitamin K (10mg) was minor (TP 40% after 6 hours).
What it is your opinion about indication to transfuse FFP in this case to correct prothrombin complex?
It would be sufficient reserves liver cells that respond to the vitamin K?
Thank you in advance for your help.
I AM AGRE, ONLY İF BLEEDİNG AND DICFollowing
- Sultan Salah added an answer:Can diabetic people donate blood?Blood is an all-important life saving component that any person can voluntarily donate and for which demand is steep. However, conflicting reports and suggestions by the medical fraternity indicate that willing diabetics are often deprived of the privilege of blood donation. Please clarify this point.
There is a certainty that the Type 1 DM patients are excluded from donating blood even though they are keen to donate, as regards to the type 2 DM they can donate subject to they full fill the criteria (for instance additional co-morbidities like CV diseases,high blood pressure,metabolic syndrome etc.,) set forth by the respective health authorities of the countries, nowadays there is stringent guidelines in place for most of the countries and is readily available for the donors in the official web site of the federal health authorities.
I suppose that i have added a point to the discussion.Following
- Could red blood cells, which were removed from patients with Polycythemia vera, be used for blood transfusion (without any infectious diseases)? Phlebotomy or bloodletting has been the mainstay of therapy for the polycythemia vera (PV) disease process for a long time. The object is to remove excess cellular elements, mainly red blood cells, to improve the circulation of blood by lowering the blood viscosity. (these words were cited from http://emedicine.medscape.com/article/205114-treatment).
After red blood cells were removed from patients with PV, could they be used for blood transfusion (without any infectious diseases)?Dear José,
Your answer perfectly summarizes the strategy.
- What is the scientific background in your country for the proposed transfusion trigger for RBC? Who proposes a transfusion trigger for packed red blood cell concentrates (RBC) in your country? The medical association? Specific medical specialities like anaesthesiologists, surgeons, transfusion medicine specialists, etc.? What is the scientific basis (e.g. publications, clinical study results, etc.) for this trigger? Is it mandatory to follow this recommendation? Who controls this? Is there any penalty for not adhering to the proposed trigger?Dear Romi,
Thanks a lot.
This is helpful. You are quite advanced "down under" also in PBM - congratulations!
- Farooq Wani added an answer:Does SOD decrease or increase in Thalassemic patients?A Thalassemia patient is chronic and transfusion dependent, and as a result, they have iron overload in their bodies. I have read many journals concerning SOD activity and this iron overload. However, some researchers found it will increase SOD, and the others say decreased. I read their explanation, but it's not enough to get deep explanation based on a biochemical reason. Any answers will be appreciated.SOD activity will increase to fight increased oxidative stress inside the cellsFollowing
- Can hypertension patients donate allogenic blood? In Spain, since 2005, "severe hypertension" is a "definitive exclusion criteria". But, which is the rational severe hypertension definition? Must we accept them? And when must we reject them?
It is well known that donors under HTA treatment can have severe hypotension reaction after whole blood donation and the incidence of hypotension transfusion reactions in patients transfused with plasma from HTA donors is well known.
Thank you very much for your help, comments and experiences.Thanks
At Europe, we ussualy do not accept new donors older than 60, but veterans can continue donating after 65 y/o till 70.
They are not oldies now, only mature!Following
- Michele Schiavulli added an answer:Does anyone have a validated 'Knowledge, Attitudes and Practices (KAP) survey on blood donations?I'm seeking to carry out a survey in my home country and would prefer a validated questionnaire. I've seen a couple mentioned in papers but am unable to find them electronically. Thanks for your help.Sorry, here is the questionnaire attachment....Following
- Valencia Nkambule added an answer:Can we create universal ABO recipients like AB blood group by introducing antigens in individuals in intrauterine life?I think that in intrauterine life, if we introduce antigens at the time of development of immune system, the body will consider them as self antigens and will not produce antibodies later in life as happens in case of AB blood group. If it is possible,we can save many lives that are wasted due to ABO incompatibility.I do not believe that this will work (unless done through molecular genetics procedure) and even if it were it were, this would also diminish the donor supply during this transition period (as we are creating universal recipients). Simply introducing the antigen in embryonic life will cause the babies to be born with the antibodies instead, and the reason that this is not is the fact that when babies are born they have to prior exposure to the antigens, However they start to build them up as early as 3 months of age.Following
- Why do all we still transfuse two Red Blood Cells Concentrates each time? ASH’s Chosing Wisely first recommendation advises against liberal transfusion of RBCs:
"Transfusion of the smallest effective dose of RBCs is recommended because, compared with restrictive strategies, liberal transfusion does not improve patient outcomes.
Therefore, liberal transfusion generates costs and exposes patients to potential harms from transfusion without likelihood of benefit."
Consistent with this recommendation, ASH panel further advise that clinicians avoid administering 2 units of RBCs if 1 unit is sufficient and that appropriate weight-based dosing of RBCs be used in children.Hi Ivo!
Actually at my hospital we are serving, except in case of active bleeding, the unit one by one. Also, one today and tomorrow "maybe" the second one, after reevaluation!
Also, after review all the "transfusion order" and "hemoglobin level", I try to phone to each prescritor. I had small service presentation in the most important consume service: "treat the patient, not the figure!
Also, in the hemogram study we are treating to add a message like: We recommend the anemia study, or give iron, etc; plus, we recommend the apply of restrictive transfusion criteria: one by one, ajusted to clinical situation and acording cardiovascular risk factors!
We have reduced more than 30%the red cell concentrates in three years. Also we have save more than half millon of euros last years!
But the problem are the own hematologist! My main "enemy" is my own new boss!!
We must fight every day again pre 1988 NIH Conference attitudes!!
I hope to see at Oporto (NATA Meeting)Following
- Mohammad I Khan added an answer:Blood transfusions in mice?We have a mutant line of mice in which the homozygous mutant pups die around P7 from hemolytic anemia (the mutated protein is important in red blood cells). We'd like to study the mutant gene/protein's function in the brain, and for this we need to get the pups to survive a few weeks longer. We'd like to try blood transfusions (from mom to pup) but I can't find much info out there about doing this in mice.
Specifically, my questions are:
1. Do you need to wash/purify mom's blood before injecting it into the pup?
2. Should I collect blood into an anticoagulant? If so, which one won't affect the recipient pup?
3. Is blood-typing necessary in mice?As long as your work has nothing to do with coagulation, use any one you like. Sodium citrate is easy and cheapest. I have used sodium citrate and worked well.Following
- Khaled Z added an answer:What is the best practice to optimize preoperative Hb?What is your practice, recommendations, and experience in optimizing preoperative Hb? When do you give Iron and Erythropoietin? Which one is superior? Any other alternatives to minimize perioperative blood transfusion?Dear Abdullah,
Optimizing preoperative Hb, depends on the subject, the reason of the anemia, and more on the expected blood loss during surgery. I agree with M Mueller for at least 3 weeks for workout; in France we can give IV Iron (if iron deficiency objectively diagnosed only in hospitals and clinics since january 2014 ) or Blood ptransfusion if urgent. EPO still for refractory anemia or renal insufficiency.Following
- Seyed-Reza Mortazavi added an answer:How much plasma can be removed from whole blood?Is there any formula or tables to predict the amount of plasma that can be removed using volume, hematocrit and the centrifugation settings? I give you an example, I'd like to know in advance how much plasma I can remove after centrifugating a 600 cc whole blood bag with 7% hematocrit during 15 minutes at 200g(rcf) at room temperature.There is no unique formula for this purpose. The amount of maximum plasma volume to be collected is related to the volume and Hct of the original blood unit, the Hct of the resultant packed red cells, whether or not the collection of plasma has therapeutic purposes (i.e. it is plasmapheresis), and other factors.Following
- Maria Baou added an answer:Plasmapheresis-whole blood transfusion and myeloma?In the light that bortezomib causes peripheral neuropathy, do you think that plasmapheresis during bortezomib treatment followed by whole blood transfusion (not replacement of patient's WBC, RBC) will improve adverse effects of bortezomib and response to treatment in multiple myeloma patients?In fact I believe that after the end of any kind of chemotherapy (not only Bortezomib) all Leukemia patients should receive up to 500 ml of fresh blood from healthy volunteers to replenish the anemia, neutropenia, pancytopenia and myelosuppression caused by the chemotherapy. I also believe that frequent blood transfusion after the end of chemotherapy (once a month) would be more beneficial to the patients than stem cell transplantation because healthy plasma from normal donors will provide Immunoglobulins, complement, cloting factors etc resulting in fewer infections and less bleeding in the vast majority of Leukemia types treated with standard chemotherapeutic agents. Besides circulating stem cells or messenchymal stem cells or circulating endothelial progenitor cells that would be in blood from healthy volunteers would initiate myelopoiesis and angiogenesis (for damaged from chemotherapy endothelial cells and capillaries) much faster and possibly more efficiently than stem cell transplantation especially if we take under consideration the side effects of immunosuppressants.Following
- Maria Baou asked a question:CXCR4 mobilisation and blood transfusion in CLL?Do you think that following mobilization of CLL cells from peripheral organs (with anti-CXCR4 or other drugs), whole blood apheresis followed by whole blood transfusion before antibody treatment (Rituximab, anti-CD19) will help responses to antibody treatment in CLL?Following
About Blood Transfusion
The introduction of whole blood or blood component directly into the blood stream. (Dorland, 27th ed)